Single-shot super-resolution total internal reflection fluorescence microscopy

Guo, Min; Chandris, Panagiotis; Giannini, John; Trexler, Adam J.; Fischer, R.; Chen, Jiji; Vishwasrao, Harshad D.; Rey-Suarez, Ivan; Wu, Yicong; Wu, Xufeng; Waterman, Clare M.; Patterson, George H.; Upadhyaya, Arpita; Taraska, Justin W.; Shroff, Hari

doi:10.1038/s41592-018-0004-4

Cited by 62 publications

(36 citation statements)

References 38 publications

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“…The high-resolution information from SIM images can be extracted from Fourier-transformed images in a process called deconvolution. Several illumination modes have been used to create SIM excitation patterns, e.g., in 3D [45], in a total internal reflection mode [46,47] at a grazing incidence angle [48] or in a lattice light sheet [49]. The final SIM images show a lateral resolution of typically 100-130 nm and an axial resolution of 100-250 nm that depends on the wavelength of emission light, the numerical aperture (NA) of the objective, the distance from the coverslip, and the illumination mode.…”

Section: Structured Illumination Microscopy (Sim)mentioning

confidence: 99%

Super-Resolution Imaging of Tight and Adherens Junctions: Challenges and Open Questions

Gonschior

Haucke

Lehmann

2020

IJMS

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The tight junction (TJ) and the adherens junction (AJ) bridge the paracellular cleft of epithelial and endothelial cells. In addition to their role as protective barriers against bacteria and their toxins they maintain ion homeostasis, cell polarity, and mechano-sensing. Their functional loss leads to pathological changes such as tissue inflammation, ion imbalance, and cancer. To better understand the consequences of such malfunctions, the junctional nanoarchitecture is of great importance since it remains so far largely unresolved, mainly because of major difficulties in dynamically imaging these structures at sufficient resolution and with molecular precision. The rapid development of super-resolution imaging techniques ranging from structured illumination microscopy (SIM), stimulated emission depletion (STED) microscopy, and single molecule localization microscopy (SMLM) has now enabled molecular imaging of biological specimens from cells to tissues with nanometer resolution. Here we summarize these techniques and their application to the dissection of the nanoscale molecular architecture of TJs and AJs. We propose that super-resolution imaging together with advances in genome engineering and functional analyses approaches will create a leap in our understanding of the composition, assembly, and function of TJs and AJs at the nanoscale and, thereby, enable a mechanistic understanding of their dysfunction in disease.

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Section: Structured Illumination Microscopy (Sim)mentioning

confidence: 99%

Super-Resolution Imaging of Tight and Adherens Junctions: Challenges and Open Questions

Gonschior

Haucke

Lehmann

2020

IJMS

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“…Among these SRMs, SIM is suitable for fast live-cell imaging and has been used to reveal numerous subcellular structures and dynamics (Li et al, 2015;Nixon-Abell et al, 2016;Zhanghao et al, 2019). Recently, ER-PM MCS in U2OS, Jurkat T and HEK293 cells was observed by SIM with total internal reflection fluorescence microscopy (TIRF-SIM) (Guo M. et al, 2018;Kang et al, 2019). The grazing incidence SIM (GI-SIM) was developed and applied to visualize ER-mitochondria, ER-late endosome and ER-lysosome MCSs in COS-7 and U2OS cells (Guo Y. et al, 2018).…”

Section: Visualization Of Mcss By Em and Srmmentioning

confidence: 99%

Current and Emerging Approaches for Studying Inter-Organelle Membrane Contact Sites

Huang

Jiang

et al. 2020

Front. Cell Dev. Biol.

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Inter-organelle membrane contact sites (MCSs) are classically defined as areas of close proximity between heterologous membranes and established by specific proteins (termed tethers). The interest on MCSs has rapidly increased in the last years, since MCSs play a crucial role in the transfer of cellular components between different organelles and have been involved in important cellular functions such as apoptosis, organelle division and biogenesis, and cell growth. Recently, an unprecedented depth and breadth in insights into the details of MCSs have been uncovered. On one hand, extensive MCSs (organelles interactome) are revealed by comprehensive analysis of organelle network with high temporal-spatial resolution at the system level. On the other hand, more and more tethers involving in MCSs are identified and further works are focusing on addressing the role of these tethers in regulating the function of MCSs at the molecular level. These enormous progresses largely depend on the powerful approaches, including several different types of microscopies and various biochemical techniques. These approaches have greatly accelerated recent advances in MCSs at the system and molecular level. In this review, we summarize the current and emerging approaches for studying MCSs, such as various microscopies, proximity-driven fluorescent signal generation and proximity-dependent biotinylation. In addition, we highlight the advantages and disadvantages of the techniques to provide a general guidance for the study of MCSs.

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“…For example, SIM has been applied to study spreading behavior and changes in cytoskeletal rearrangement in platelets with cytoskeletal protein deficiency; such as ARPC1-deficiency, where reduced actinrelated protein 2/3 complex (Arp2/3) led to aberrant platelet spreading (66) and Wiskott-Aldrich syndrome protein (WASp) deficient platelets from patients and WASp knockout mice resulted in reduced actin nodule formation (65). With the expanding implementation of SIM in other microscopy methods (e.g., TIRF microscopy), we anticipate SIM to have increased applications in high to super-resolution imaging of platelet and thrombus behavior (101).…”

Section: Nanoscale Imaging Of Single Plateletsmentioning

confidence: 99%

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

et al. 2020

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Platelets are small anucleate cells that are essential for many biological processes including hemostasis, thrombosis, inflammation, innate immunity, tumor metastasis, and wound healing. Platelets circulate in the blood and in order to perform all of their biological roles, platelets must be able to arrest their movement at an appropriate site and time. Our knowledge of how platelets achieve this has expanded as our ability to visualize and quantify discreet platelet events has improved. Platelets are exquisitely sensitive to changes in blood flow parameters and so the visualization of rapid intricate platelet processes under conditions found in flowing blood provides a substantial challenge to the platelet imaging field. The platelet's size (∼2 µm), rapid activation (milliseconds), and unsuitability for genetic manipulation, means that appropriate imaging tools are limited. However, with the application of modern imaging systems to study platelet function, our understanding of molecular events mediating platelet adhesion from a single-cell perspective, to platelet recruitment and activation, leading to thrombus (clot) formation has expanded dramatically. This review will discuss current platelet imaging techniques in vitro and in vivo, describing how the advancements in imaging have helped answer/expand on platelet biology with a particular focus on hemostasis. We will focus on platelet aggregation and thrombus formation, and how platelet imaging has enhanced our understanding of key events, highlighting the knowledge gained through the application of imaging modalities to experimental models in vitro and in vivo. Furthermore, we will review the limitations of current imaging techniques, and questions in thrombosis research that remain to be addressed. Finally, we will speculate how the same imaging advancements might be applied to the imaging of other vascular cell biological functions and visualization of dynamic cell-cell interactions.

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Single-shot super-resolution total internal reflection fluorescence microscopy

Cited by 62 publications

References 38 publications

Super-Resolution Imaging of Tight and Adherens Junctions: Challenges and Open Questions

Super-Resolution Imaging of Tight and Adherens Junctions: Challenges and Open Questions

Current and Emerging Approaches for Studying Inter-Organelle Membrane Contact Sites

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

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